橡胶树硫氧还蛋白基因HbCXXS1的克隆及表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2022-0290

摘要/Abstract

摘要：

硫氧还蛋白通过调控细胞内的氧化还原状态在植物生长发育及非生物胁迫应答等过程中发挥重要作用。研究硫氧还蛋白基因HbCXXS1在橡胶树各组织以及激素和非生物胁迫处理下的表达，为解析其生物学功能奠定基础。采用RT-PCR克隆HbCXXS1，对其进行生物信息学分析，并采用实时荧光定量PCR分析该基因在橡胶树不同组织及不同激素和非生物胁迫条件下的表达模式。结果表明，HbCXXS1开放阅读框为372 bp，编码123个氨基酸。HbCXXS1蛋白为亲水性蛋白，具有硫氧还蛋白保守结构域，活性中心为CXXS，属于h型硫氧还蛋白的第III组。HbCXXS1在橡胶树胶乳中的表达显著高于其他组织，且在死皮植株胶乳中的表达显著低于健康植株。脱落酸、茉莉酸甲酯、干旱和盐胁迫处理抑制了HbCXXS1的表达，乙烯利和低温处理上调了HbCXXS1的表达，而水杨酸和过氧化氢处理的不同时间点，HbCXXS1的表达既有上调也有下调。HbCXXS1可能在橡胶树产排胶和非生物胁迫应答中发挥重要调控作用。

关键词: 硫氧还蛋白, 橡胶树, 克隆, 表达分析, 非生物胁迫

Abstract:

Thioredoxin plays an important role in plant growth and development and abiotic stress response by regulating cellular redox homeostasis. This study aims to investigate the expressions of thioredoxin gene HbCXXS1 in various tissues of rubber tree（Hevea brasiliensis）and under different hormones and abiotic stresses，for laying a foundation for analyzing its biological function. The HbCXXS1 gene was cloned by RT-PCR and analyzed by bioinformatics. Quantitative real-time PCR was applied to analyze the expression patterns of HbCXXS1 in different tissues of rubber tree and in response to various hormones and abiotic stresses. The results showed that HbCXXS1 contained an open reading frame of 372 bp encoding 123 amino acids. HbCXXS1 was a hydrophilic protein，and contained a conserved thioredoxin domain with a CXXS active site，which belonged to group III of thioredoxin h. HbCXXS1 in the latex was expressed significantly higher than that in other tissues. In addition，its expression was significantly decreased in latex of tapping panel dryness trees as compared with that of healthy trees. The expression of HbCXXS1 was inhibited by abscisic acid，methyl jasmonate，drought and salt stresses but was up-regulated by ethephon and cold stress. Moreover，HbCXXS1 expression was both up-regulated and down-regulated at different time points after salicylic acid and hydrogen peroxide treatments. These results suggested that HbCXXS1 might play an important regulatory role in latex production and latex flow，as well as abiotic stress responses in rubber tree.

Key words: thioredoxin, rubber tree, cloning, expression analysis, abiotic stress

王帅, 袁坤, 何其光, 胡义钰, 冯成天, 王真辉, 刘进平, 刘辉. 橡胶树硫氧还蛋白基因HbCXXS1的克隆及表达分析[J]. 生物技术通报, 2022, 38(12): 214-222.

WANG Shuai, YUAN Kun, HE Qi-guang, HU Yi-yu, FENG Cheng-tian, WANG Zhen-hui, LIU Jin-ping, LIU Hui. Cloning and Expression Analysis of HbCXXS1，a Thioredoxin Gene in Hevea brasiliensis[J]. Biotechnology Bulletin, 2022, 38(12): 214-222.

图/表 6

图1 HbCXXS1的PCR扩增

Fig. 1 PCR amplification of HbCXXS1 gene

图2 HbCXXS1蛋白保守结构域（A）、跨膜结构域（B）和亲/疏水性（C）分析

Fig. 2 Conserved domain（A），transmembrane domain（B）and hydrophilicity/hydrophobicity（C）analysis of HbCXXS1 protein

图3 HbCXXS1和其他植物h型硫氧还蛋白的系统进化树

Fig. 3 Phylogenetic tree of HbCXXS1 and thioredoxin h proteins in other plants

图4 HbCXXS1在橡胶树各组织（A）以及健康与死皮植株胶乳（B）中的表达不同小写字母表示差异显著（P<0.05）。下同

Fig. 4 Expressions of HbCXXS1 in various tissues（A）and latex from healthy and TPD rubber tree（B） Different lowercase letters indicate a significant difference（P<0.05）. The same below

图5 激素处理后HbCXXS1的表达变化

Fig. 5 Expression change of HbCXXS1 after hormone treatment

图6 非生物胁迫下HbCXXS1的表达变化

Fig. 6 Expression change of HbCXXS1 under abiotic stress

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